Response to androgen therapy in patients with dyskeratosis congenita.
Dyskeratosis congenita (DC) is an inherited bone marrow failure syndrome and telomere biology disorder characterized by dysplastic nails, reticular skin pigmentation and oral leucoplakia. Androgens are a standard therapeutic option for bone marrow failure in those patients with DC who are unable to undergo haematopoietic stem cell transplantation, but there are no systematic data on its use in those patients. We evaluated haematological response and side effects of androgen therapy in 16 patients with DC in our observational cohort study. Untreated DC patients served as controls. Seventy percent of treated DC patients had a haematological response with red blood cell and/or platelet transfusion independence. The expected age-related decline in telomere length was noted in androgen-treated patients. All treated DC patients had at least one significant lipid abnormality. Additional treatment-related findings included a significant decrease in thyroid binding globulin, accelerated growth in pre-pubertal children and splenic peliosis in two patients. Liver enzymes were elevated in both androgen-treated and untreated patients, suggesting underlying liver involvement in DC. This study suggests that androgen therapy can be effectively used to treat bone marrow failure in DC, but that side effects need to be closely monitored.


Congenital myasthenic syndromes.
Congenital myasthenic syndromes (CMSs) are a genotypically and phenotypically heterogeneous group of neuromuscular disorders, which have in common an impaired neuromuscular transmission. Since the field of CMSs is steadily expanding, the present review aimed at summarizing and discussing current knowledge and recent advances concerning the etiology, clinical presentation, diagnosis, and treatment of CMSs. Systematic literature review. Currently, mutations in 32 genes are made responsible for autosomal dominant or autosomal recessive CMSs. These mutations concern 8 presynaptic, 4 synaptic, 15 post-synaptic, and 5 glycosilation proteins. These proteins function as ion-channels, enzymes, or structural, signalling, sensor, or transporter proteins. The most common causative genes are CHAT, COLQ, RAPSN, CHRNE, DOK7, and GFPT1. Phenotypically, these mutations manifest as abnormal fatigability or permanent or fluctuating weakness of extra-ocular, facial, bulbar, axial, respiratory, or limb muscles, hypotonia, or developmental delay. Cognitive disability, dysmorphism, neuropathy, or epilepsy are rare. Low- or high-frequency repetitive nerve stimulation may show an abnormal increment or decrement, and SF-EMG an increased jitter or blockings. Most CMSs respond favourably to acetylcholine-esterase inhibitors, 3,4-diamino-pyridine, salbutamol, albuterol, ephedrine, fluoxetine, or atracurium. CMSs are an increasingly recognised group of genetically transmitted defects, which usually respond favorably to drugs enhancing the neuromuscular transmission. CMSs need to be differentiated from neuromuscular disorders due to muscle or nerve dysfunction.


Mixed plantations enhance more soil organic carbon stocks than monocultures across China: Implication for optimizing afforestation/reforestation strategies.
Forests play an essential role in mitigating climate change by sequestering carbon dioxide from the atmosphere. The establishment of mixed plantations is a promising way to store carbon (C) in soil compared with monocultures. However, monoculture forests largely dominate the rapid increase in forest areas in China. To optimize afforestation strategies and maximize the subsequent potential of C sequestration, we conducted a meta-analysis with 427 observations across 176 study sites in China. The goal was to quantify changes in the stocks of soil organic carbon (SOC) in mixed plantations compared with monocultures and to identify the predominant drivers for the stocks of SOC, including geological location, climatic factors, land use history, edaphic properties, plantation age, the inclusion of nitrogen-fixing trees, mixing proportion, and mixed plant types. The results showed that mixed plantations significantly increased the SOC stocks by 12% compared with monocultures, and the mixing proportion should not exceed 55% to produce higher SOC stocks in mixed plantations compared with monoculture. Additionally, mixed plantations in barren land are the most likely to increase the SOC stocks with limited water or low temperatures for growth. Additional measures instead of mixed plantations should be explored to increase SOC stocks in north, central, and northwest China. The data from this study demonstrated the spatiotemporal variability on the storage of SOC driven by mixed trees and has valuable implications for the establishment and management of afforestation.


Tracking effluent discharges in undisturbed stony soil and alluvial gravel aquifer using synthetic DNA tracers.
With the intensification of human activities, fresh water resources are increasingly being exposed to contamination from effluent disposal to land. Thus, there is a greater need to identify the sources and pathways of water contamination to enable the development of better mitigation strategies. To track discharges of domestic effluent into soil and groundwater, 10 synthetic double-stranded DNA (dsDNA)3 tracers were developed in this study. Laboratory column experiment and field groundwater and soil lysimeter studies were carried out spiking DNA with oxidation-pond domestic effluent. The selected DNA tracers were compared with a non-reactive bromide (Br) tracer with respect to their relative mass recoveries, speeds of travel and dispersions using the method of temporal moments. In intact stony soil and gravel aquifer media, the dsDNA tracers typically showed earlier breakthrough and less dispersion than the Br tracer, and underwent mass reduction. This suggests that the dsDNA tracers were predominantly transported through the network of larger pores or preferential flow paths. Effluent tracking experiments in soil and groundwater demonstrated that the dsDNA tracers were readily detectable in effluent-contaminated soil and groundwater using quantitative polymerase chain reaction. DNA tracer spiked in the effluent at quantities of 36μg was detected in groundwater 37m down-gradient at a concentration 3-orders of magnitude above the detection limit. It is anticipated it could be detected at far greater distances. Our findings suggest that synthetic dsDNA tracers are promising for tracking effluent discharges in soils and groundwater but further studies are needed to investigate DNA-effluent interaction and the impact of subsurface environmental conditions on DNA attenuation. With further validation, synthetic dsDNA tracers, especially when multiple DNA tracers are used concurrently, can be an effective new tool to track effluent discharge in soils and groundwater, providing spatial estimation on the presence or absence of contamination sources and pathways.


Sparse patch-based label propagation for accurate prostate localization in CT images.
In this paper, we propose a new prostate computed tomography (CT) segmentation method for image guided radiation therapy. The main contributions of our method lie in the following aspects. 1) Instead of using voxel intensity information alone, patch-based representation in the discriminative feature space with logistic sparse LASSO is used as anatomical signature to deal with low contrast problem in prostate CT images. 2) Based on the proposed patch-based signature, a new multi-atlases label fusion method formulated under sparse representation framework is designed to segment prostate in the new treatment images, with guidance from the previous segmented images of the same patient. This method estimates the prostate likelihood of each voxel in the new treatment image from its nearby candidate voxels in the previous segmented images, based on the nonlocal mean principle and sparsity constraint. 3) A hierarchical labeling strategy is further designed to perform label fusion, where voxels with high confidence are first labeled for providing useful context information in the same image for aiding the labeling of the remaining voxels. 4) An online update mechanism is finally adopted to progressively collect more patient-specific information from newly segmented treatment images of the same patient, for adaptive and more accurate segmentation. The proposed method has been extensively evaluated on a prostate CT image database consisting of 24 patients where each patient has more than 10 treatment images, and further compared with several state-of-the-art prostate CT segmentation algorithms using various evaluation metrics. Experimental results demonstrate that the proposed method consistently achieves higher segmentation accuracy than any other methods under comparison.


Systemic organ wasting induced by localized expression of the secreted insulin/IGF antagonist ImpL2.
Organ wasting, related to changes in nutrition and metabolic activity of cells and tissues, is observed under conditions of starvation and in the context of diseases, including cancers. We have developed a model for organ wasting in adult Drosophila, whereby overproliferation induced by activation of Yorkie, the Yap1 oncogene ortholog, in intestinal stem cells leads to wasting of the ovary, fat body, and muscle. These organ-wasting phenotypes are associated with a reduction in systemic insulin/IGF signaling due to increased expression of the secreted insulin/IGF antagonist ImpL2 from the overproliferating gut. Strikingly, expression of rate-limiting glycolytic enzymes and central components of the insulin/IGF pathway is upregulated with activation of Yorkie in the gut, which may provide a mechanism for this overproliferating tissue to evade the effect of ImpL2. Altogether, our study provides insights into the mechanisms underlying organ-wasting phenotypes in Drosophila and how overproliferating tissues adapt to global changes in metabolism.


Methods to calculate the heat index as an exposure metric in environmental health research.
Environmental health research employs a variety of metrics to measure heat exposure, both to directly study the health effects of outdoor temperature and to control for temperature in studies of other environmental exposures, including air pollution. To measure heat exposure, environmental health studies often use heat index, which incorporates both air temperature and moisture. However, the method of calculating heat index varies across environmental studies, which could mean that studies using different algorithms to calculate heat index may not be comparable. We investigated 21 separate heat index algorithms found in the literature to determine a) whether different algorithms generate heat index values that are consistent with the theoretical concepts of apparent temperature and b) whether different algorithms generate similar heat index values. Although environmental studies differ in how they calculate heat index values, most studies' heat index algorithms generate values consistent with apparent temperature. Additionally, most different algorithms generate closely correlated heat index values. However, a few algorithms are potentially problematic, especially in certain weather conditions (e.g., very low relative humidity, cold weather). To aid environmental health researchers, we have created open-source software in R to calculate the heat index using the U.S. National Weather Service's algorithm. We identified 21 separate heat index algorithms used in environmental research. Our analysis demonstrated that methods to calculate heat index are inconsistent across studies. Careful choice of a heat index algorithm can help ensure reproducible and consistent environmental health research.


Search for resonant top-antitop production in the lepton plus jets decay mode using the full CDF data set.
This Letter reports a search for a narrow resonant state decaying into two W bosons and two b quarks where one W boson decays leptonically and the other decays into a quark-antiquark pair. The search is particularly sensitive to top-antitop resonant production. We use the full data sample of proton-antiproton collisions at a center-of-mass energy of 1.96 TeV collected by the CDF II detector at the Fermilab Tevatron, corresponding to an integrated luminosity of 9.45 fb(-1). No evidence for resonant production is found, and upper limits on the production cross section times branching ratio for a narrow resonant state are extracted. Within a specific benchmark model, we exclude a Z' boson with mass, M(Z'), below 915 GeV/c(2) decaying into a top-antitop pair at the 95% credibility level assuming a Z' boson decay width of Γ(Z') = 0.012 M(Z'). This is the most sensitive search for a narrow qq-initiated tt resonance in the mass region below 750 GeV/c(2).


Availability and Accuracy of Information Regarding Nonprescription Emergency Contraception.
Although access to emergency contraception (EC) has increased with nonprescription status and approval of Plan B One-Step without age restrictions, barriers may still remain in patient education. This study assesses product availability and accuracy of information for EC among community pharmacies in Rhode Island, comparing changes from 2009 to 2012. Two female investigators posing as patients seeking EC followed a standardized script over telephone conversations. Investigators assessed EC availability, product use information, and cost at all community (retail) pharmacies in Rhode Island. Data were reported as group results with no identifiers. Chi-square and Fisher exact tests were used to analyze results. During spring of 2009 and 2012, 165 and 171 pharmacies were telephoned, respectively. Approximately 90% of pharmacies stocked EC both years. In all, 62% versus 28% (P < .001) indicated EC should be taken as soon as possible; 82.5% versus 87.7% (P = .220) provided correct administration information; 67% versus 84% (P < .001) warned about adverse effects; and 67% versus 53% (P = .123) provided the correct minimum age for purchase. Access to nonprescription EC in Rhode Island is very good. Sites not stocking EC should reassess plans for patients to obtain medication. There is need for reeducation on EC labeling to improve counseling provided over the telephone.


Utilization of dental services in old age.
Regular utilization of dental services helps to improve and maintain oral and general health, even in old age. However, utilization behavior for dental services changes with age: preventive utilization behavior is often replaced by a symptom-driven one. With age, a decrease in the utilization of dental services can be observed, whilst the frequency of contact with physicians increases. The present review describes the current knowledge regarding the utilization of dental services in old age (frequency, reasons for non-utilization, modifying factors). The reasons for non-utilization of dental services are multidimensional: subjective reasons and other objective modifying factors can be distinguished. The frequency of utilization also differs with personal context and attitude. On the basis of the available evidence no conclusive explanation could be provided. A checklist should allow dental practitioners to monitor the factors that affect the utilization of services within their own dental office.


Improving anterior deltoid activity in a musculoskeletal shoulder model - an analysis of the torque-feasible space at the sternoclavicular joint.
Modelling the shoulder's musculature is challenging given its mechanical and geometric complexity. The use of the ideal fibre model to represent a muscle's line of action cannot always faithfully represent the mechanical effect of each muscle, leading to considerable differences between model-estimated and in vivo measured muscle activity. While the musculo-tendon force coordination problem has been extensively analysed in terms of the cost function, only few works have investigated the existence and sensitivity of solutions to fibre topology. The goal of this paper is to present an analysis of the solution set using the concepts of torque-feasible space (TFS) and wrench-feasible space (WFS) from cable-driven robotics. A shoulder model is presented and a simple musculo-tendon force coordination problem is defined. The ideal fibre model for representing muscles is reviewed and the TFS and WFS are defined, leading to the necessary and sufficient conditions for the existence of a solution. The shoulder model's TFS is analysed to explain the lack of anterior deltoid (DLTa) activity. Based on the analysis, a modification of the model's muscle fibre geometry is proposed. The performance with and without the modification is assessed by solving the musculo-tendon force coordination problem for quasi-static abduction in the scapular plane. After the proposed modification, the DLTa reaches 20% of activation.


Plasma membrane proteomics of human breast cancer cell lines identifies potential targets for breast cancer diagnosis and treatment.
The use of broad spectrum chemotherapeutic agents to treat breast cancer results in substantial and debilitating side effects, necessitating the development of targeted therapies to limit tumor proliferation and prevent metastasis. In recent years, the list of approved targeted therapies has expanded, and it includes both monoclonal antibodies and small molecule inhibitors that interfere with key proteins involved in the uncontrolled growth and migration of cancer cells. The targeting of plasma membrane proteins has been most successful to date, and this is reflected in the large representation of these proteins as targets of newer therapies. In view of these facts, experiments were designed to investigate the plasma membrane proteome of a variety of human breast cancer cell lines representing hormone-responsive, ErbB2 over-expressing and triple negative cell types, as well as a benign control. Plasma membranes were isolated by using an aqueous two-phase system, and the resulting proteins were subjected to mass spectrometry analysis. Overall, each of the cell lines expressed some unique proteins, and a number of proteins were expressed in multiple cell lines, but in patterns that did not always follow traditional clinical definitions of breast cancer type. From our data, it can be deduced that most cancer cells possess multiple strategies to promote uncontrolled growth, reflected in aberrant expression of tyrosine kinases, cellular adhesion molecules, and structural proteins. Our data set provides a very rich and complex picture of plasma membrane proteins present on breast cancer cells, and the sorting and categorizing of this data provides interesting insights into the biology, classification, and potential treatment of this prevalent and debilitating disease.


Coronavirus S protein-induced fusion is blocked prior to hemifusion by Abl kinase inhibitors.
Enveloped viruses gain entry into host cells by fusing with cellular membranes, a step that is required for virus replication. Coronaviruses, including the severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV) and infectious bronchitis virus (IBV), fuse at the plasma membrane or use receptor-mediated endocytosis and fuse with endosomes, depending on the cell or tissue type. The virus spike (S) protein mediates fusion with the host cell membrane. We have shown previously that an Abelson (Abl) kinase inhibitor, imatinib, significantly reduces SARS-CoV and MERS-CoV viral titres and prevents endosomal entry by HIV SARS S and MERS S pseudotyped virions. SARS-CoV and MERS-CoV are classified as BSL-3 viruses, which makes experimentation into the cellular mechanisms involved in infection more challenging. Here, we use IBV, a BSL-2 virus, as a model for studying the role of Abl kinase activity during coronavirus infection. We found that imatinib and two specific Abl kinase inhibitors, GNF2 and GNF5, reduce IBV titres by blocking the first round of virus infection. Additionally, all three drugs prevented IBV S-induced syncytia formation prior to the hemifusion step. Our results indicate that membrane fusion (both virus-cell and cell-cell) is blocked in the presence of Abl kinase inhibitors. Studying the effects of Abl kinase inhibitors on IBV will be useful in identifying the host cell pathways required for coronavirus infection. This will provide an insight into possible therapeutic targets to treat infections by current as well as newly emerging coronaviruses.


Block copolymers as bile salt sequestrants: intriguing structures formed in a mixture of an oppositely charged amphiphilic block copolymer and bile salt.
To study the formation and characterize the structure of mixed complexes of oppositely charged block copolymers and surfactants are of great significance for practical applications, e.g., in drug carrier formulations that are based on electrostatically assisted assembly. In this context, biocompatible block copolymers and biosurfactants (like bile salts) are particularly interesting. In this work, we report on the co-assembly in dilute aqueous solution between a cationic poly(N-isopropyl acryl amide) (PNIPAM) diblock copolymer and the oppositely charged bile salt surfactant sodium deoxycholate at ambient temperature. The cryogenic transmission electron microscopy (cryo-TEM) experiments revealed the co-existence of two types of co-assembled complexes of radically different morphology and inner structure. They are formed mainly as a result of the electrostatic attraction between the positively charged copolymer blocks and bile salt anions and highlight the potential of using linear amphiphilic block copolymers as bile salt sequestrants in the treatment of bile acid malabsorption and hypercholesterolemia. The first complex of globular morphology has a coacervate core of deoxycholate anions and charged copolymer blocks surrounded by a PNIPAM corona. The second complex has an intriguing tape-like supramolecular morphology of several micrometer in length that is striped in the direction of the long axis. A model is presented in which the stretched cationic blocks of several block copolymers interact electrostatically with the bile salt molecules that are associated to form a zipper-like structure. The tape is covered on both sides by the PNIPAM chains that stabilize the overall complex in solution. In addition to cryo-TEM, the mixed system was investigated in a range of molar charge fractions at a constant copolymer concentration by static light scattering, small angle X-ray scattering, and electrophoretic mobility measurements.


The impact of longstanding illness and common mental disorder on competing employment exits routes in older working age: A longitudinal data-linkage study in Sweden.
Comorbidity is prevalent in older working ages and might affect employment exits. This study aimed to 1) assess the associations between comorbidity and different employment exit routes, and 2) examine such associations by gender. We used data from employed adults aged 50-62 in the Stockholm Public Health Survey 2002 and 2006, linked to longitudinal administrative income records (N = 10,416). The morbidity measure combined Limiting Longstanding Illness and Common Mental Disorder-captured by the General Health Questionnaire-12 (≥4)-into a categorical variable: 1) No Limiting Longstanding Illness, no Common Mental Disorder, 2) Limiting Longstanding Illness only, 3) Common Mental Disorder only, and 4) comorbid Limiting Longstanding Illness+Common Mental Disorder. Employment status was followed up until 2010, treating early retirement, disability pension and unemployment as employment exits. Competing risk regression analysed the associations between morbidity and employment exit routes, stratifying by gender. Compared to No Limiting Longstanding Illness, no Common Mental Disorder, comorbid Limiting Longstanding Illness+Common Mental Disorder was associated with early retirement in men (subdistribution hazard ratio = 1.73, 95% confidence intervals: 1.08-2.76), but not in women. For men and women, strong associations for disability pension were observed with Limiting Longstanding Illness only (subdistribution hazard ratio = 11.43, 95% confidence intervals: 9.40-13.89) and Limiting Longstanding Illness+Common Mental Disorder (subdistribution hazard ratio = 14.25, 95% confidence intervals: 10.91-18.61), and to a lesser extent Common Mental Disorder only (subdistribution hazard ratio = 2.00, 95% confidence intervals: 1.31-3.05). Women were more likely to exit through disability pension than men (subdistribution hazard ratio = 1.96, 95% confidence intervals: 1.60-2.39). Common Mental Disorder only was the only morbidity category associated with unemployment (subdistribution hazard ratio = 1.70, 95% confidence intervals: 1.36-2.15). Strong associations were observed between specific morbidity categories with different employment exit routes, which differed by gender. Initiatives to extend working lives should consider older workers' varied health needs to prevent inequalities in older age.


Transplantation of umbilical cord mesenchymal stem cells alleviates lupus nephritis in MRL/lpr mice.
Systemic lupus erythematosus (SLE) is a multisystem autoimmune disease, which, despite the advances in immunosuppressive medical therapies, remains potentially fatal in some patients, especially in treatment-refractory patients. This study found that transplantation of umbilical cord mesenchymal stem cells (UC-MSCs) has the same therapeutic effect as transplantation of bone marrow mesenchymal stem cells (BM-MSCs), which has been reported to be efficient in treating SLE-related symptoms in MRL/lpr mice. Multi-treatment (at the 18th, 19th, and 20th weeks of age) of 1 × 10(6) UC-MSCs was able to decrease the levels of 24-h proteinuria, serum creatinine, and anti-double-stranded DNA (dsDNA) antibody, and the extent of renal injury such as crescent formation in MRL/lpr mice. A lower, but still significant, reduction in these parameters was also observed in mice receiving a single dose of UC-MSCs (at the 18th week). UC-MSCs treatment also inhibited expression of monocyte chemotactic protein-1 (MCP-1) and high-mobility group box 1 (HMGB-1) expression in a similar fashion. UC-MSCs labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE) were found in the lungs and kidneys 1 week post infusion. In addition, after 11 weeks post UC-MSCs infusion, human cells were found in kidney of UC-MSCs-treated mice. These findings indicated that UC-MSCs transplantation might be a potentially promising approach in the treatment of lupus nephritis, possibly by inhibiting MCP-1 and HMGB-1 production.


Acidosis-induced metallothionein (MT) mRNA expression in neonatal rat primary astrocyte cultures.
Metallothionein (MT) mRNA levels were determined following exposure of neonatal rat primary astrocyte cultures to acidosis. Astrocyte total RNA was probed on northern blots with [alpha 32 P]dCTP-labeled synthetic cDNA probes specific for rat MT isoform mRNAs. The probe for MT-I mRNA hybridized to a single mRNA with a size appropriate for MT, approximately 550 nucleotides. MT-I mRNA levels in astrocyte monolayers exposed to pH 6.5 and 6.0 for 3 or 6 hours were unchanged compared with MT-I mRNA levels in control cultures exposed to pH 7.4. In contrast, 9 hour exposure of astrocytes to pH 6.5 and 6.0 led to a significant increase in MT-I mRNA transcripts compared with controls maintained at pH 7.4 (p < 0.001 and p < 0.02, respectively). A probe for MT-II mRNA that hybridizes to a single mRNA (450 nucleotides) was also used to determine the effect of acidosis on astrocyte MT-II mRNA transcripts. Although statistical significance was not attained, a similar trend was noted, with a 9 hour exposure to pH of 6.5 and 6.0 resulting in increased astrocytic expression MT-II mRNA compared with control cells maintained at pH 7.4. Acidosis was also associated with a pH-dependent increase in astrocytic volume. Accordingly, acidosis is invoked as an added stimulus to stress factors associated with the induction of astrocytic MT mRNA transcripts.


Extensive genomic diversity among Mycobacterium marinum strains revealed by whole genome sequencing.
Mycobacterium marinum is the causative agent for the tuberculosis-like disease mycobacteriosis in fish and skin lesions in humans. Ubiquitous in its geographical distribution, M. marinum is known to occupy diverse fish as hosts. However, information about its genomic diversity is limited. Here, we provide the genome sequences for 15 M. marinum strains isolated from infected humans and fish. Comparative genomic analysis of these and four available genomes of the M. marinum strains M, E11, MB2 and Europe reveal high genomic diversity among the strains, leading to the conclusion that M. marinum should be divided into two different clusters, the "M"- and the "Aronson"-type. We suggest that these two clusters should be considered to represent two M. marinum subspecies. Our data also show that the M. marinum pan-genome for both groups is open and expanding and we provide data showing high number of mutational hotspots in M. marinum relative to other mycobacteria such as Mycobacterium tuberculosis. This high genomic diversity might be related to the ability of M. marinum to occupy different ecological niches.


Heterogeneity of the genome ancestry of individuals classified as White in the state of Rio Grande do Sul, Brazil.
One hundred nineteen individuals classified as White, living in different localities of the Brazilian state of Rio Grande do Sul, were studied in relation to the HVS-I region of the mitochondrial DNA (mtDNA). The male fraction of the sample (N = 74) was also tested for seven Y-chromosome polymorphisms. In a specific population (Veranópolis), a city characterized by a large influence of the Italian immigration of the 19th century, the results from the maternal and paternal sides indicated almost complete European ancestry. However, another sample identified as White, from different localities of Rio Grande do Sul, presented significant fractions of Native American (36%) and African (16%) mtDNA haplogroups. These results indicate that Brazilian populations are remarkably heterogeneous; while some present an overwhelming majority of transplanted European genomes, with a complete correspondence between physical appearance and ancestry, others reflect a history of extensive admixture with dissociation between physical appearance and ancestry.


RNA-dependent DNA polymerase activity of RNA tumor virus. VI. Processive mode of action of avian myeloblastosis virus polymerase.
Purified avian myeloblastosis virus (AMV) polymerase consisting of alpha,beta subunits has been shown to act processively in catalyzing DNA synthesis primed with 34S AMV RNA oligo(dT), poly(A)-poly(dT), and poly(I)-poly(dC). DNA transcripts prepared with 34S AMV RNA-oligo(dT)14 and AMV polymerase (alphabeta) have been shown to have a molecular weight of 1.05 X 10(6), or approximately one-third the size of the 34S RNA genome. Polymerase subunit alpha acts nonprocessively with the above templates.


Disability as an equal opportunity issue within nurse education in the UK.
Whilst equal opportunity policies, related research and subsequent curricula changes over recent years may have begun to have a positive impact in some areas of nursing education and practice, 'disability' struggles for recognition as an equality issue. Disabled people, making up approximately six million of the UK population, experience discrimination as part of their daily lives and this is also evident in the nursing services they receive. Far from challenging discrimination, nursing education may simply sustain the negative dominant ideological view of disability. Drawing comparisons with 'race', which has an established equal opportunity dimension, this paper explores disability equality, which is relatively neglected in nursing curricula. From this review it is argued that nursing as a profession has a role beyond patient care in influencing public perceptions concerning disabled people and should take more positive steps in promoting social justice by challenging discrimination. Stakeholders in nursing curricula and nursing practice have both legal and professional responsibilities to ensure that nursing and nurses become part of the solution to disability discrimination. Including disability in equal opportunity policies and strengthening their impact on pre and post registration nursing curricula, provides nursing with a tangible opportunity to do just that.


A novel micropit device integrates automated cell positioning by dielectrophoresis and nuclear transfer by electrofusion.
Nuclear transfer (NT) cloning involves manual positioning of individual donor-recipient cell couplets for electrofusion. This is time-consuming and introduces operator-dependent variation as a confounding parameter in cloning trials. In order to automate the NT procedure, we developed a micro-fluidic device that integrates automated cell positioning and electrofusion of isolated cell couplets. A simple two layer micro-fluidic device was fabricated. Thin film interdigitated titanium electrodes (300 nm thick, 250 microm wide and 250 microm apart) were deposited on a solid borosilicate glass substrate. They were coated with a film of electrically insulating photosensitive epoxy polymer (SU-8) of either 4 or 22 microm thickness. Circular holes ("micropits") measuring 10, 20, 30, 40 or 80 microm in diameter were fabricated above the electrodes. The device was immersed in hypo-osmolar fusion buffer and manually loaded with somatic donor cells and recipient oocytes. Dielectrophoresis (DEP) was used to attract cells towards the micropit and form couplets on the same side of the insulating film. Fusion pulses between 80 V and 120 V were applied to each couplet and fusion scored under a stereomicroscope. Automated couplet formation between oocytes and somatic cells was achieved using DEP. Bovine oocyte-oocyte, oocyte-follicular cells and oocyte-fibroblast couplets fused with up to 69% (n = 13), 50% (n = 30) and 78% (n = 9) efficiency, respectively. Fusion rates were comparable to parallel plate or film electrodes that are conventionally used for bovine NT. This demonstrates proof-of-principle that a micropit device is capable of both rapid cell positioning and fusion.


Evolution of the endomembrane systems of trypanosomatids - conservation and specialisation.
Parasite surfaces support multiple functions required for survival within their hosts, and maintenance and functionality of the surface depends on membrane trafficking. To understand the evolutionary history of trypanosomatid trafficking, where multiple lifestyles and mechanisms of host interactions are known, we examined protein families central to defining intracellular compartments and mediating transport, namely Rabs, SNAREs and RabGAPs, across all available Euglenozoa genomes. Bodonids possess a large trafficking repertoire, which is mainly retained by the Trypanosoma cruzi group, with extensive losses in other lineages, particularly African trypanosomes and phytomonads. There are no large-scale expansions or contractions from an inferred ancestor, excluding direct associations between parasitism or host range. However, we observe stepwise secondary losses within Rab and SNARE cohorts (but not RabGAPs). Major changes are associated with endosomal and late exocytic pathways, consistent with the diversity in surface proteomes between trypanosomatids and mechanisms of interaction with the host. Along with the conserved core family proteins, several lineage-specific members of the Rab (but not SNARE) family were found. Significantly, testing predictions of SNARE complex composition by proteomics confirms generalised retention of function across eukaryotes.


Differential effects of serum heat treatment on chemotaxis and phagocytosis by human neutrophils.
Neutrophils, in cooperation with serum, are vital gatekeepers of a host's microbiome and frontline defenders against invading microbes. Yet because human neutrophils are not amenable to many biological techniques, the mechanisms governing their immunological functions remain poorly understood. We here combine state-of-the-art single-cell experiments with flow cytometry to examine how temperature-dependent heat treatment of serum affects human neutrophil interactions with "target" particles of the fungal model zymosan. Assessing separately both the chemotactic as well as the phagocytic neutrophil responses to zymosan, we find that serum heat treatment modulates these responses in a differential manner. Whereas serum treatment at 52°C impairs almost all chemotactic activity and reduces cell-target adhesion, neutrophils still readily engulf target particles that are maneuvered into contact with the cell surface under the same conditions. Higher serum-treatment temperatures gradually suppress phagocytosis even after enforced cell-target contact. Using fluorescent staining, we correlate the observed cell behavior with the amounts of C3b and IgG deposited on the zymosan surface in sera treated at the respective temperatures. This comparison not only affirms the critical role of complement in chemotactic and adhesive neutrophil interactions with fungal surfaces, but also unmasks an important participation of IgGs in the phagocytosis of yeast-like fungal particles. In summary, this study presents new insight into fundamental immune mechanisms, including the chemotactic recruitment of immune cells, the adhesive capacity of cell-surface receptors, the role of IgGs in fungal recognition, and the opsonin-dependent phagocytosis morphology of human neutrophils. Moreover, we show how, by fine-tuning the heat treatment of serum, one can selectively study chemotaxis or phagocytosis under otherwise identical conditions. These results not only refine our understanding of a widely used laboratory method, they also establish a basis for new applications of this method.


Single-dose metronidazole clears Opalina sp. from juvenile Bufo woodhousii.
Protozoans of the family Opalinidae are intestinal commensals in amphibians. To test the hypothesis that these organisms are susceptible to the antiprotozoal antibiotic metronidazole, we randomly assigned 60 juvenile Woodhouse's toads ( Bufo woodhousii ) to receive a single oral dose of metronidazole or water. In pilot trials, the prevalence of opalinids in untreated members of this population was over 70%. One-third of the study population was dissected at each of 3 time points: 18 hr, 1 wk, and 2 wk post-treatment. An examiner blinded to the toad's treatment history determined the presence or absence of opalinids using a dissecting microscope. Opalinids were found in 3/10 toads in the treatment group and 9/10 in the control group after 18 hr (P < 0.02), in none of the treatment group and 8/10 in the control group after 1 wk (P < 0.001), and in none of the treatment group and 10/10 in the control group after 2 wk (P < 0.0001). These results suggest that a single-dose of metronidazole quickly and reliably clears opalinids from juvenile Woodhouse's toads with no evidence of short-term recurrence. The treatment was well tolerated, with no apparent morbidity and no mortality in either group. Future exploration of opalinid-related host fitness consequences may be facilitated by this simple method of developing a protozoan-free host population.


Outcomes from a national multispecialty palliative care curriculum development project.
In 1998 we completed a successful regional pilot project in palliative care curriculum development among 32 internal medicine residency programs recruited from the mid-western United States. Between 1999 and 2004 this project was expanded to include 358 U.S. programs, from four specialties, based on new training requirements in internal medicine, family medicine, neurology, and general surgery. To assess the 1-year outcomes from residency programs participating in a national multispecialty palliative care curriculum development project. Outcome data obtained from residency programs' responses to a structured progress report 12 months after enrolling in the project and from published residency project reports. Three hundred fifty-eight residency programs, representing 27% of all eligible training programs in the four specialties, participated in the project. Outcome data was available from 224 residencies (63%). Most programs started new teaching in pain, non-pain symptom management, and communication skills. More than 50% of programs integrated palliative care topics within established institutional grand rounds, morbidity/mortality conferences or morning report. More than 70% of internal medicine and family practice programs began new direct patient care training opportunities utilizing hospital-based palliative care or hospice programs. New faculty development initiatives and use of quality improvement projects to drive curriculum change were reported in less than 50% of programs. Focused short-term instruction in palliative care curriculum development, in a diverse group of residency programs, is feasible and associated with significant curriculum change.


A liquid crystalline precursor incorporating chlorhexidine acetate and silver nanoparticles for root canal disinfection.
Lyotropic liquid crystals (LLC) have received increasing attention as a drug delivery system. In this study, a novel intra-canal disinfectant based on the glycerol monooleate (GMO) LLC precursor incorporation with chlorhexidine (CHX) and silver nanoparticles (Ag-NPs) was designed and evaluated. The LLC precursor with excellent fluidity was able to penetrate deeply into the complex tiny collateral branch root canals. The transformation of cubic LLC in root canals upon coming into contact with water provided long-lasting disinfection against multidrug-resistant bacteria to avoid the endodontic reinfection and follow-up visits. The GMO-ethanol-water (48% : 12% : 40%, w/w) formulation containing 0.5% CHX and 0.02% Ag-NPs was selected for further studies. The low viscosity of the precursor presented excellent injectability and flowabilities. From the in vitro release test, the release behaviours were found to be influenced by CHX and Ag-NP contents, allowing the optimized precursor to obtain a 28-day release profile. The CHX-Ag-NP containing LLC precursor exhibited an excellent and sustained sterilization effect on Enterococcus faecalis for more than one month with a bacterial inactivation rate of ≥98.5%, which was far more than the minimum clinical requirement (7 days). Furthermore, no in vitro toxicity was observed in the cytotoxicity evaluation. The CHX-Ag-NP containing LLC precursor was proved to be a promising intra-canal disinfectant in our study.


Cell interactions in nerve and muscle cell cultures.
The neurotransmitter synthesized by a given class of neurones is subject to modification and, indeed, a qualitative switchover in transmitter biochemistry recently has been demonstrated (Furshpan, POtter & Landis, 1980; Walicke, Campenot & Patterson, 1977). In conjunction with the specification of transmitter biosynthesis that becomes established in a given neurone, a complementary specification of appropriate receptor production is required in any cell functionally post-synaptic to that neurone. An additional requirement of peculiar force in the nervous system has to do with the spatial organization of the receptors in the surface membrane of the post-synaptic cell once the receptors are synthesized. Inappropriately distributed receptors are useless receptors. The perfect registration of a variety of types of presynaptic release sites with high post-synaptic concentrations of appropriate receptors constitutes one of the outstanding features of nervous-system organization that must be accounted for. We report some experiments directed toward understanding the cell biology of regulation of receptor distribution over the surface membrane of muscle cells. Functional synaptic connexions are formed quite early in development and the stability and maturation of synaptic networks is contingent on a number of factors. One interesting contingency is that related to the functional activity of developing networks. Do only those networks survive and mature which are activated by stimuli impinging from the environment? (Wiesel & Hubel, 1963). Put more simple, are action potentials and synaptic activity essential for neuronal maturation? We address this question in cell culture systems from the mammalian central nervous system.


Investigating the structural plasticity of a cytochrome P450: three-dimensional structures of P450 EryK and binding to its physiological substrate.
Cytochrome P450s are heme-containing proteins that catalyze the oxidative metabolism of many physiological endogenous compounds. Because of their unique oxygen chemistry and their key role in drug and xenobiotic metabolism, particular attention has been devoted in elucidating their mechanism of substrate recognition. In this work, we analyzed the three-dimensional structures of a monomeric cytochrome P450 from Saccharopolyspora erythraea, commonly called EryK, and the binding kinetics to its physiological ligand, erythromycin D. Three different structures of EryK were obtained: two ligand-free forms and one in complex with its substrate. Analysis of the substrate-bound structure revealed the key structural determinants involved in substrate recognition and selectivity. Interestingly, the ligand-free structures of EryK suggested that the protein may explore an open and a closed conformation in the absence of substrate. In an effort to validate this hypothesis and to investigate the energetics between such alternative conformations, we performed stopped-flow absorbance experiments. Data demonstrated that EryK binds erythromycin D via a mechanism involving at least two steps. Contrary to previously characterized cytochrome P450s, analysis of double jump mixing experiments confirmed that this complex scenario arises from a pre-existing equilibrium between the open and closed subpopulations of EryK, rather than from an induced-fit type mechanism.


Overcoming endocrine resistance due to reduced PTEN levels in estrogen receptor-positive breast cancer by co-targeting mammalian target of rapamycin, protein kinase B, or mitogen-activated protein kinase kinase.
Activation of the phosphatidylinositol 3-kinase (PI3K) pathway in estrogen receptor α (ER)-positive breast cancer is associated with reduced ER expression and activity, luminal B subtype, and poor outcome. Phosphatase and tensin homolog (PTEN), a negative regulator of this pathway, is typically lost in ER-negative breast cancer. We set out to clarify the role of reduced PTEN levels in endocrine resistance, and to explore the combination of newly developed PI3K downstream kinase inhibitors to overcome this resistance. Altered cellular signaling, gene expression, and endocrine sensitivity were determined in inducible PTEN-knockdown ER-positive/human epidermal growth factor receptor 2 (HER2)-negative breast cancer cell and/or xenograft models. Single or two-agent combinations of kinase inhibitors were examined to improve endocrine therapy. Moderate PTEN reduction was sufficient to enhance PI3K signaling, generate a gene signature associated with the luminal B subtype of breast cancer, and cause endocrine resistance in vitro and in vivo. The mammalian target of rapamycin (mTOR), protein kinase B (AKT), or mitogen-activated protein kinase kinase (MEK) inhibitors, alone or in combination, improved endocrine therapy, but the efficacy varied by PTEN levels, type of endocrine therapy, and the specific inhibitor(s). A single-agent AKT inhibitor combined with fulvestrant conferred superior efficacy in overcoming resistance, inducing apoptosis and tumor regression. Moderate reduction in PTEN, without complete loss, can activate the PI3K pathway to cause endocrine resistance in ER-positive breast cancer, which can be overcome by combining endocrine therapy with inhibitors of the PI3K pathway. Our data suggests that the ER degrader fulvestrant, to block both ligand-dependent and -independent ER signaling, combined with an AKT inhibitor is an effective strategy to test in patients.


[Special aspects of pediatric anesthesia in ophthalmic surgery].
In ophthalmic surgery, children are anesthetized for various reasons. The airway needs special attention as children have higher oxygen requirements and brief periods of apnea can result in hypoxemia and subsequent bradycardia or even cardiac arrest. Most interventions can be managed with the aid of a laryngeal mask allowing minor airway manipulations and thereby reducing the risk of laryngeal and tracheal injuries and laryngospasm in children with hypersensitive airways. In children older than 3 years an adequate postoperative nausea and vomiting (PONV) prophylaxis should be performed. The modified postoperative vomiting in children (POVOC) score is a suitable method for the estimation of PONV. Strabismus surgery is an independent risk factor for PONV; therefore, prophylaxis is recommended in any case. The combination of drugs from different substance groups amplifies the antiemetic effect. This article discusses whether anesthesia can damage immature brains. In animal experiments it could be proven that neuronal apoptosis could be induced by most of the commonly used anesthetics. It has not yet been clarified whether this has an effect on the neurocognitive development of children. Because of this uncertainty the indications for surgery and anesthesia should be assessed on strict clinical requirements. There is, however, widespread agreement that a necessary anesthesia carried out in a correct and controlled manner has no negative consequences for children.


Eyelid and scleral necrosis following bare iridium-192 seed for retinoblastoma.
A 2 1/2-year-old girl with a history of bilateral retinoblastoma underwent primary enucleation of the right eye and was referred for further management of persistent tumor in the fellow eye. Previous treatment of the left eye included external beam radiotherapy, systemic chemotherapy, laser photocoagulation, cryotherapy, and direct scleral application of a bare iridium-192 radioactive seed. Examination revealed focal full-thickness necrosis of the left upper and lower eyelid and a large inferonasal viable retinoblastoma tumor with overlying retinal detachment. Systemic chemotherapy and direct laser photocoagulation were administered. Four months after presentation, the patient developed focal scleral necrosis with 360 degrees hemorrhagic choroidal detachment. Enucleation was performed and histopathologic examination demonstrated full-thickness scleral necrosis with adjacent viable retinoblastoma tumor cells. Follow-up examinations showed no evidence of recurrent or metastatic tumor. This case is the first report of scleral necrosis following combined modality treatment of retinoblastoma.


Immunological characteristics of hyperreactive malarial splenomegaly syndrome in sudanese patients.
Hyperreactive Malarial Splenomegaly (HMS) is defined as a massive enlargement of the spleen resulting from abnormal immune responses after repeated exposure to the malaria parasites. This study was carried out in Khartoum, Sudan. Sudan is considered to be one of the countries where HMS is quite prevalent. The objective of the study was to determine the incidence of HMS in patients who reported to the Omdurman Tropical Diseases Hospital (OMTDH) in Sudan and to investigate the basic laboratory and immunological characteristics of this condition in these patients. A cross-sectional study was carried out in OMTDH, and all patients with enlarged spleens were included in the study. Thirty-one out of 335 (9.3%) patients were diagnosed as having the HMS condition using international criteria for HMS diagnosis. The mean serum immunoglobulin M (IgM) levels in HMS patient groups were 14.3 ± 5 g/L, and this was significantly higher compared with geographically matched controls (P < 0.001). Immunoglobulin G (IgG) C anticircumsporozoite (CSP) antibody levels were higher in the HMS patients although the difference was not statistically significant, when compared with a group of patients with mild malaria. In comparison with naïve European controls, both the HMS and the mild malaria groups had significantly higher antimalarial antibody levels P < 0.001 and P < 0.01, respectively. Plasma levels of interleukin 10 (IL10) and interferon gamma (IFN γ ) were significantly increased in the HMS patients compared with the healthy control donors (P < 0.05 and P < 0.01) for IL10 and IFN γ , respectively. The findings of this study suggest that HMS is one of the significant causes of tropical splenomegaly in Sudan. HMS is associated with significant elevations of circulating IgM and antimalarial IgG antibodies as well as IL10 and IFN γ .


Nosological profile in a dermatology referral center in the state of Amazonas -Brazil.
Fundaments: Skin diseases are associated wih high morbidity, low mortality and low rate of hospitalization. However, they can cause considerable interference in physical and emotional well-being of the individual. Several of them reach large population, requiring specific interventions for their control. To describe the frequency of skin disease diagnosed in the dermatology service in Manaus, capital of Amazonas State. We collected data on registered sex, age, origin and diagnostics for the first consultation of patients attended between January 2000 and December 2007. Of the 56.024 recorded visits, we obtained 56.720 cases of dermatological diagnoses, being the most common sexually transmitted diseases (25,12%), allergic skin disesases (14,03%), unspecified dermatoses (13,01%), leprosy (6,34%) and acne, seborrhea and related diseases (5,05%). The frequency was similar for both sexes, aged 20-29 years predominated and Manaus the origin most reported. The pattern of skin diseases identified in this study may serve as a baseline to managers of health system in the region develop strategies for prevention and control of dermatoses, with emphasis on sexually transmitted diseases, allergic skin diseases, leprosy and acne.


Analysis of endocannabinoid signaling elements and related proteins in lymphocytes of patients with Dravet syndrome.
Cannabidiol (CBD) reduces seizures in childhood epilepsy syndromes including Dravet syndrome (DS). A formulation of CBD has obtained orphan drug designation for these syndromes and clinical trials are currently underway. The mechanism responsible for CBD effects is not known, although it could involve targets sensitive to CBD in other neurological disorders. We believe of interest to investigate whether these potential targets are altered in DS, in particular whether the endocannabinoid system is dysregulated. To this end, lymphocytes from patients and controls were used for analysis of gene expression of transmitter receptors and transporters, ion channels, and enzymes associated with CBD effects, as well as endocannabinoid genes. Plasma endocannabinoid levels were also analyzed. There were no differences between DS patients and controls in most of the CBD targets analyzed, except an increase in the voltage-dependent calcium channel α-1h subunit. We also found that cannabinoid type-2 (CB 2) receptor gene expression was elevated in DS patients, with no changes in other endocannabinoid-related receptors and enzymes, as well as in plasma levels of endocannabinoids. Such elevation was paralleled by an increase in CD70, a marker of lymphocyte activation, and certain trends in inflammation-related proteins (e.g., peroxisome proliferator-activated receptor-γ receptors, cytokines). In conclusion, together with changes in the voltage-dependent calcium channel α-1h subunit, we found an upregulation of CB 2 receptors, associated with an activation of lymphocytes and changes in inflammation-related genes, in DS patients. Such changes were also reported in inflammatory disorders and may indirectly support the occurrence of a potential dysregulation of the endocannabinoid system in the brain.


Biodegradation of haloacetic acids by bacterial isolates and enrichment cultures from drinking water systems.
Biodegradation is a potentially important loss process for haloacetic acids (HAAs), a class of chlorination byproducts, in water treatment and distribution systems, but little is known about the organisms involved (i.e., identity, substrate range, biodegradation kinetics). In this research, 10 biomass samples (i.e., tap water, distribution system biofilms, and prechlorinated granular activated carbon filters) from nine drinking water systems were used to inoculate a total of thirty enrichment cultures fed monochloroacetic acid (MCAA), dichloroacetic acid (DCAA), or trichloroacetic (TCAA) as sole carbon and energy source. HAA degraders were successfully enriched from the biofilm samples (GAC and distribution system) but rarely from tap water. Half of the MCAA and DCAA enrichment cultures were positive, whereas only one TCAA culture was positive (two were inconclusive). Eight unique HAA-degrading isolates were obtained including several Afipia spp. and a Methylobacterium sp.; all isolates were members of the phylum Proteobacteria. MCAA, monobromoacetic acid (MBAA), and monoiodoacetic acid (MIAA) were rapidly degraded by all isolates, and DCAA and tribromoacetic (TBAA) were also relatively labile. TCAA and dibromoacetic acid (DBAA)were degraded by only three isolates and degradation lagged behind the other HAAs. Detailed DCAA biodegradation kinetics were obtained for two selected isolates and two enrichment cultures. The maximum biomass-normalized degradation rates (Vm) were 0.27 and 0.97 microg DCAA/ microg protein/h for Methylobacterium fujisawaense strain PAWDI and Afipia felis strain EMD2, respectively, which were comparable to the values obtained for the enrichment cultures from which those organisms were isolated (0.39 and 1.37 microg DCAN/microg protein/h, respectively). The half-saturation constant (Km) values ranged from 4.38 to 77.91 microg DCAA/L and the cell yields ranged from 14.4 to 36.1 mg protein/g DCAA.


Initial stages of tumor cell-induced angiogenesis: evaluation via skin window chambers in rodent models.
There is a paucity of information about events that follow immediately after tumor cells are triggered to initiate the process of angiogenesis (the formation of new blood vessels). Such information is relevant to the issue of when micrometastases vascularize and has implications for the accessibility of micrometastases to various treatments. In this study, we attempted to monitor events at the initiation of angiogenesis at the earliest possible stage of tumor growth in vivo. Two different rodent mammary tumor cell lines, R3230Ac from the Fischer 344 rat and 4T1 from the BALB/c mouse, were stably transfected with a gene that encodes an enhanced version of green fluorescence protein (GFP). GFP-labeled R3230Ac or 4T1 cells (about 20-50 cells) were implanted into dorsal skinfold window chambers of Fischer 344 rats or BALB/c mice, respectively. Tumor angiogenesis was then monitored serially and noninvasively for up to 4 weeks. Clear evidence of modification of the host vasculature was observed when tumor mass reached approximately 60-80 cells, and functional new blood vessels were seen when tumor mass reached roughly 100-300 cells. Individual tumor cells exhibited a chemotaxis-like growth pattern toward the pre-existing host vasculature. When ex-flk1 (a soluble, truncated vascular endothelial cell growth factor receptor protein known to be antiangiogenic) was injected with the tumor cells, the initial angiogenic and tumor growth activities were inhibited considerably, indicating that angiogenesis inhibitors may halt tumor growth even before the onset of angiogenesis. Angiogenesis induced by tumor cells after implantation in the host begins at a very early stage, i.e., when the tumor mass contains roughly 100-300 cells. Identification of chemotactic signals that initiate tumor cell migration toward the existing vasculature may provide valuable targets for preventing tumor progression and/or metastases.


Programmable fiber-based picosecond optical pulse shaper using time-domain binary phase-only linear filtering.
We demonstrate a fiber-based programmable arbitrary picosecond optical pulse shaper using binary phase-only linear filtering. The reconfigurable filtering operation is implemented in the time domain using an electro-optical phase modulator driven by a high-speed bit pattern generator. The required binary phase code is designed using a genetic algorithm. Precise matching between the predispersive and postdispersive media in the system is achieved by use of a single linearly chirped fiber Bragg grating subsequently operated from its two input ends. As a proof of concept, different pulse waveforms of practical interest are generated using this new pulse shaper.


Altered subunit communication in subfamilies of trimeric dUTPases.
The enzyme dUTPase is essential in preventing uracil incorporation into DNA. Design of antagonists against this novel chemotherapeutic target requires identification of species-specific differences in the structure and mechanism of the enzyme. This task is now approached via comparisons of available crystallographic structures of dUTPases from Homo sapiens, Escherichia coli, and retroviruses. The eukaryotic protein uniquely displays polar and charged amino acid residues participating in threefold intersubunit interactions. In bacterial and retroviral dUTPases, threefold interactions are mainly hydrophobic. The residues responsible for this contrast are mapped in multiple sequence alignment to positions differently and characteristically conserved in distinct evolutionary branches. The general feature of this contrast is further strengthened by a second eukaryotic model structure constructed using comparative modeling. The dUTPase cDNA from Drosophila melanogaster was identified, sequenced, and the model structure of the encoded polypeptide displayed a polar hydrogen-bonding network of threefold interactions, identically to the human structure. Results allow clear distinction between two subfamilies of trimeric dUTPases where altered subunit communication may account for a functional difference in the catalytic cycle.


Simple tone discriminations are disrupted following experimental frontal traumatic brain injury in rats.
To assess cognitive deficits in a rat model of brain injury. Cognitive deficits are some of the most pervasive and enduring symptoms of frontal traumatic brain injury (TBI) in human patients. In animal models, the assessment of cognitive deficits from TBI has primarily been limited to tests of spatial learning. Recently, simple discrimination performance has been shown to be sensitive to frontal brain damage. The current study provides a detailed characterization of deficits in a two-choice tone discrimination following a bilateral frontal controlled cortical impact injury. Rats were trained on a two-tone discrimination task in a standard operant chamber, then either a frontal brain injury was delivered or sham procedures performed. Following recovery, they were re-tested on the discrimination task and then tested on a reversal of the discrimination. Frontal injury caused substantial deficits in responding and discrimination accuracy as well as an increase in side bias. Based on the outcomes seen in this study, discrimination and other operant tasks may provide a sensitive tool to assess the effect of therapeutic agents on cognitive deficits in animal models, which could lead to improved characterization of deficits and yield an improved assessment tool to aid in drug discovery.


Combined active and passive heat exposure induced heat acclimation in a soccer referee before 2014 FIFA World Cup.
The 2014 FIFA World Cup was held in Brazil, where the climatic conditions presented a significant thermoregulatory and perceptual challenge to those unfamiliar with the heat and humidity. This case report documents the adaptation induced by a novel mixed methods (isothermic and passive) heat acclimation (HA) regime for a northern European professional soccer match official prior to the tournament. The intervention involved 13 HA sessions over an 18 day period comprising five isothermic HA sessions whereby intermittent running was used to target and maintain tympanic temperature (Tytemp) at 38 °C for 90 min, and seven passive HA sessions of 48 °C water bathing for 30 min. The athlete performed a heat stress test (HST) (35 min running at four incremental intensities in 30 °C) and a repeated high-intensity running test (as many 30 s self-paced efforts as possible, to a maximum of 20, with 30 s passive recovery) before and after the intervention. The mixed methods HA regime increased plasma volume (+7.1 %), and sweat loss (+0.9 L h(-1)), reduced exercising Tytemp (-0.6 °C), and mean body temperature (-0.5 °C). High-intensity running performance improved after HA (+29 %), as did the perception of thermal comfort during exercise (-0.3 units). This data evidences the effectiveness of a practical, mixed methods HA strategy, remotely implemented around training and competition, at inducing the heat acclimation phenotype in a high-level soccer match official.


Optical properties of combustion-synthesized iron oxide aggregates.
Measurements of scattering/extinction and asymmetry ratios at both polarization orientations, along with depolarization and reciprocity ratios, corresponding to nanosized Fe2O3 aggregates were carried out in an Fe(CO)5-seeded CO air-diffusion flame as a function of the position above the burner surface. The measurements were combined with an exact light-scattering theory to yield, for the first time, the complex refractive index, the primary particle-size parameter, the aspect ratio, and the number density and volume fractions of the Fe2O3 chainlike aggregates under flame conditions. An effective complex refractive index of 1.96 +/- 6.9% to 0.20 +/- 14.5% was inferred from the data analysis at a wavelength of 488 nm and in the temperature range of 894-924 K. The corresponding primary particle size was found to be 48 mm +/- 13%, and the aggregate aspect ratio was in the range of 6-7. Comparisons of the size parameters are made with transmission electron microscopy results, and the possible sources of uncertainties in the inferred results pertaining to temperature, polydispersity effects, and the departure from the straight configuration are discussed.


Vaccination Practices in Pediatric Dialysis Patients Across Europe. A European Pediatric Dialysis Working Group and European Society for Pediatric Nephrology Dialysis Working Group Study.
Data on the immunization practices in pediatric chronic kidney disease (CKD) patients are scarce. The purpose of this study was to evaluate current vaccination practices for children on dialysis across European pediatric nephrology centers. A total of 18 tertiary pediatric nephrology centers from 12 European countries were included in the study. The data on universal national immunization programs and immunization practices for children with chronic disease or risk were recorded from European Center for Disease Prevention and Control and the World Health Organization. The immunization practices and center protocols for monitoring antibody titers after vaccination in dialysis patients were obtained through a questionnaire. All centers included in the study recommended immunization against hepatitis B virus (HBV), diphtheria, tetanus, pertussis, Hemophilus influenzae type b (Hib), poliomyelitis, measles, mumps, rubella (MMR), and streptococcus pneumonia in dialysis patients. In 16 centers, dialysis patients were vaccinated against influenza virus annually. HBV protective antibody titers were measured in 17 centers (during dialysis period in 14 centers, during pre-renal transplantation preparations in 14 centers or in both times in 11 centers). Hepatitis A virus (HAV) was reported to be followed in 13 centers, in 8 centers during dialysis period, and in 11 centers during pre-RTx preparations. MMR and varicella-zoster virus (VZV) protective antibody titers were measured during the dialysis period or before renal transplantation (RTx) in 12 and 15 centers, respectively, and in 6 centers both titers were checked both times. There are variations in vaccination practice across Europe. Children with CKD, those undergoing dialysis, and transplant candidates should receive age-appropriate vaccinations before RTx as well as before the transition to adult nephrology clinics and antibody levels should be monitored to evaluate the immunization status before and after RTx.


Measuring quality of life in oncologic breast surgery: a systematic review of patient-reported outcome measures.
Multiple randomized trials demonstrate equivalent survival between BCT and mastectomy, but clinical outcomes research must also evaluate patient satisfaction and quality of life. This review analyzes existing patient-reported outcome (PRO) measures in oncologic breast surgery to assess utility and make recommendations for future research. We performed a systematic literature review to identify PRO measures used in oncologic breast surgery patients. After applying inclusion and exclusion criteria, qualifying instruments were assessed for adherence to international guidelines for health outcomes instrument development and validation. Ten measures underwent development and psychometric evaluation in an oncologic breast surgery population. Five of ten measures (EORTC QLQ BR-23, FACT-B, HBIS, BIBCQ, and BREAST-Q) reported an adequate development and validation process. Three of these 5 measures (EORTC QLQ BR-23, FACT-B, HBIS) focused on non-surgical treatment issues. A fourth instrument (BIBCQ) did not address aesthetic concerns after breast reconstruction. The fifth instrument (BREAST-Q) was developed for use in patients undergoing mastectomy ± reconstruction, but did not address breast-conserving therapy. Overall, two key limitations were noted: 1) surgery-specific issues of breast-conserving surgery patients were not well represented and 2) measures were largely developed without the aid of newer psychometric methods that may improve their clinical utility. Reliable and valid PRO measures in breast cancer patients exist, but even the best instruments do not address all important surgery-specific and psychometric issues of oncologic breast surgery patients. Newer psychometric methods would facilitate development of scales for use in individual patient care as well as group level comparisons.


Access to knee arthroplasty among National Health Fund beneficiaries in Chile between 2004 and 2021.
Introduction: Knee osteoarthritis affects the quality of life, with knee arthroplasty being a cost-effective treatment for the severe stage of this disease. Access to knee arthroplasty is a health indicator of the Organisation for Economic Co-operation and Development. The objectives of this study are to determine the incidence of knee arthroplasty between 2004 and 2021 in beneficiaries of the National Health Fund in Chile, the proportion of patients who underwent surgery in the private system, and to estimate the patient's out-of-pocket expenditure for surgery. Methods: Cross-sectional study. We used the Department of Statistics and Health Information database. Patients discharged from a Chilean health center who underwent knee arthroplasty surgery between 2004 and 2021 were investigated. We analyzed the proportion of patients by their National Health Fund category and whether their surgery was performed in public or private network facilities. Results: Of the 31 526 knee arthroplasty procedures, 21 248 (67.38%) were performed on National Health Fund patients and 16 238 in public institutions (51.49%). Patients from the National Health Fund showed a systematic increase in knee arthroplasty volume until 2019 but decreased in 2020 and 2021 by 68% and 51%. Of the total number of patients in the public system operated on for knee arthroplasty, 856 (9%) belonged to group A1, 12 806 (60%) to group B, 2044 (10%) to group C, and 4421 (21%) to group D. The expenditure incurred by these patients was estimated to vary between 24.4% and 27.2%. The historical proportions of access to this surgery in private institutions are 7% in group A, 13% in group B, 24% in group C, and 52% in group D. Conclusion: Fifty percent of knee arthroplasty surgeries are performed in public institutions, and two-thirds are performed on patients of the National Health Fund. Forty-six percent of the C and D groups were operated in the private system. The pandemic has increased the access gap, leading to a substantial increase in the proportion of patients from the National Health Fund of the B, C, and D groups who have migrated to the private system to access this surgery.


Potential value of adenosine 5'-triphosphate (ATP) and adenosine in anaesthesia and intensive care medicine.
Extracellular adenosine and adenosine triphosphate (ATP) are involved in biological processes including neurotransmission, muscle contraction, cardiac function, platelet function, vasodilatation, signal transduction and secretion in a variety of cell types. They are released from the cytoplasm of several cell types and interact with specific purinergic receptors which are present on the surface of many cells. This review summarizes the evidence on the potential value and applicability of ATP (not restricted to ATP-MgCl(2)) and adenosine in the field of anaesthesia and intensive care medicine. It focuses, in particular, on evidence and roles in treatment of acute and chronic pain and in sepsis. Based on the evidence from animal and clinical studies performed during the last 20 years, ATP could provide a valuable addition to the therapeutic options in anaesthesia and intensive care medicine. It may have particular roles in pain management, modulation of haemodynamics and treatment of shock.


Perceptions on undertaking regular asymptomatic self-testing for COVID-19 using lateral flow tests: a qualitative study of university students and staff.
Successful implementation of asymptomatic testing programmes using lateral flow tests (LFTs) depends on several factors, including feasibility, acceptability and how people act on test results. We aimed to examine experiences of university students and staff of regular asymptomatic self-testing using LFTs, and their subsequent behaviours. A qualitative study using semistructured remote interviews and qualitative survey responses, which were analysed thematically. People who were participating in weekly testing feasibility study, between October 2020 and January 2021, at the University of Oxford. We interviewed 18 and surveyed 214 participants. Participants were motivated to regularly self-test as they wanted to know whether or not they were infected with SARS-CoV-2. Most reported that a negative test result did not change their behaviour, but it did provide them with reassurance to engage with permitted activities. In contrast, some participants reported making decisions about visiting other people because they felt reassured by a negative test result. Participants valued the training but some still doubted their ability to carry out the test. Participants were concerned about safety of attending test sites with lots of people and reported home testing was most convenient. Clear messages highlighting the benefits of regular testing for family, friends and society in identifying asymptomatic cases are needed. This should be coupled with transparent communication about the accuracy of LFTs and how to act on either a positive or negative result. Concerns about safety, convenience of testing and ability to do tests need to be addressed to ensure successful scaling up of asymptomatic testing.


Pancreas-kidney transplantation in Japan: impact of cyclosporine on the development of immunosuppressive therapy.
The number of pancreas transplants reached 16,043 worldwide in October 2001, with 1800 performed in 2000. Since the introduction of cyclosporine (CyA) in 1979, a regimen consisting of CyA, azathioprine, and steroids has been shown to improve long-term survival of clinical transplants. In Japan, the first simultaneous pancreas-kidney transplantation was performed in 1984, using organs from a brain-dead donor. This procedure was followed by 14 pancreas transplantations from cardiac arrest donors. All cases utilized a CyA-based regimen with antilymphocyte globulin or OKT3 induction. Six of the 15 recipients required less insulin postoperatively. Under a new transplant law enforced in 1997, 10 pancreas/pancreas-kidney transplantations were performed in patients diagnosed with end-stage renal failure due to diabetes mellitus type 1. In 1 patient, the graft failed due to venous thrombosis, but the other 9 recipients achieved an increased quality of life without the need for insulin or for dialysis. Pancreas transplantation represents an effective treatment worldwide and in Japan, due to the availability of CyA or tacrolimus in combination with other agents such as antilymphocyte globulin, OKT3, or mycophenolate mofetil. This investigation presents the results of pancreas-kidney transplantation in Japan, in comparison with those worldwide, and describes a recent case in Japan.


The development of race-based perceptual categorization: skin color dominates early category judgments.
Prior research on the development of race-based categorization has concluded that children understand the perceptual basis of race categories from as early as age 4 (e.g. Aboud, 1988). However, such work has rarely separated the influence of skin color from other physiognomic features considered by adults to be diagnostic of race categories. In two studies focusing on Black-White race categorization judgments in children between the ages of 4 and 9, as well as in adults, we find that categorization decisions in early childhood are determined almost entirely by attention to skin color, with attention to other physiognomic features exerting only a small influence on judgments as late as middle childhood. We further find that when skin color cues are largely eliminated from the stimuli, adults readily shift almost entirely to focus on other physiognomic features. However, 6- and 8-year-old children show only a limited ability to shift attention to facial physiognomy and so perform poorly on the task. These results demonstrate that attention to 'race' in younger children is better conceptualized as attention to skin color, inviting a reinterpretation of past work focusing on children's race-related cognition.


Antiviral treatment in schizophrenia: a randomized pilot PET study on the effects of valaciclovir on neuroinflammation.
Patients with schizophrenia experience cognitive impairment, which could be related to neuroinflammation in the hippocampus. The cause for such hippocampal inflammation is still unknown, but it has been suggested that herpes virus infection is involved. This study therefore aimed to determine whether add-on treatment of schizophrenic patients with the anti- viral drug valaciclovir would reduce hippocampal neuroinflammation and consequently improve cognitive symptoms. We performed a double-blind monocenter study in 24 male and female patients with schizophrenia, experiencing active psychotic symptoms. Patients were orally treated with the anti-viral drug valaciclovir for seven consecutive days (8 g/day). Neuroinflammation was measured with Positron Emission Tomography using the translocator protein ligand [11C]-PK11195, pre-treatment and at seven days post-treatment, as were psychotic symptoms and cognition. Valaciclovir treatment resulted in reduced TSPO binding (39%) in the hippocampus, as well as in the brainstem, frontal lobe, temporal lobe, parahippocampal gyrus, amygdala, parietal lobe, occipital lobe, insula and cingulate gyri, nucleus accumbens and thalamus (31-40%) when using binding potential (BPND) as an outcome. With total distribution volume (VT) as outcome we found essentially the same results, but associations only approached statistical significance (p = 0.050 for hippocampus). Placebo treatment did not affect neuroinflammation. No effects of valaciclovir on psychotic symptoms or cognitive functioning were found. We found a decreased TSPO binding following antiviral treatment, which could suggest a viral underpinning of neuroinflammation in psychotic patients. Whether this reduced neuroinflammation by treatment with valaciclovir has clinical implications and is specific for schizophrenia warrants further research.


Palmar Fasciitis and Polyarthritis Syndrome: A Rare Paraneoplastic Syndrome in a Patient With Prostate Carcinoma.
A 73-year-old patient was seen in our hospital for treatment of metastatic adenocarcinoma of the prostate (pT1aN0M1a R0, BRCA-2 gene mutation). Prostatectomy and regional radiotherapy were performed and goserelin, a luteinizing hormone-releasing hormone (LHRH) analog, had been started because of disease progression. Castration-resistant progressive disease developed, and enzalutamide was added. A decrease of the prostate-specific antigen (PSA) level was achieved. Before the start of enzalutamide, the patient developed bilateral pain and stiffness of both hands combined with thickening of the hands. The symptoms progressed rapidly to bilateral flexion and extension contractures. The patient became unable to tie his shoelaces and had to use adjusted cutlery to eat. Consultation of the rheumatologist, X-rays, ultrasound and palmar skin biopsy of the hands were performed. The clinical picture resembles descriptions of "palmar fasciitis and polyarthritis syndrome" (PFPAS), a rare paraneoplastic syndrome. Positive effects of immunosuppressive medication have been reported in some cases. In our patient, treatment with oral prednisone (30 mg daily) showed no effect, therefore treatment was switched to methylprednisone pulses and methotrexate. PFPAS is an uncommon paraneoplastic syndrome characterized by rapid onset of bilateral arthritis of the hands, fasciitis of the palms, progressive stiffness and contractures. The scarcity of knowledge about PFPAS makes it difficult to recognize it at an early stage. As a paraneoplastic syndrome, it has been linked to various malignancies. Thus far, PFPAS has been described in only two other cases of prostate cancer.


Association of fibronectin and vinculin with focal contacts and stress fibers in stationary hamster fibroblasts.
We have recently observed a transmembrane association between extracellular fibronectin (FN) fibers and elongated focal patches or fibers of vinculin (VN) in G1-arrested stationary Nil 8 hamster fibroblasts, with double-label immunofluorescence microscopy (Singer and Paradiso, 1981, Cell. 24:481-492). We hypothesized that these FN-VN complexes might correspond to focal contacts, the membrane sites that are probably mainly responsible for attaching cells to their substrata, because vinculin is often localized in focal contacts. However, because fibronectin-vinculin associations may not be restricted to the substrate adhesive surface of the cell, it became necessary to determine whether some or all of the various kinds of FN-VN complexes which we described are in proximity to the substrate. Using interference reflection optics and double-label immunofluorescence microscopy for fibronectin and vinculin, many elongated (up to 38 micrometer) FN-VN associations were found to be strikingly coincident with focal contacts in the perinuclear area of extremely flattened arrested Nil 8 fibroblasts in 0.3% fetal bovine serum (FBS). In addition, the long FN-VN adhesion complexes were precisely aligned with the major phase-dense stress fibers observed at the ventral surfaces of these stationary cells with phase contrast microscopy. Fibronectin was neither associated with vinculin-containing focal contacts of Nil 8 cells cultured in medium with 5% FBS nor with vinculin-negative focal contacts located at the extreme edges of stationary cells arrested in 0.3 FBS. Our time-course experiments suggest that early FN-VN lacking-focal contacts, which form at the cellular margins, develop into mature substrate adhesion complexes containing both fibronectin and vinculin, localized in the major stress fibers at the centers of sessile fibroblasts.


Prediction of preschool aggression from DRD4 risk, parental ADHD symptoms, and home chaos.
This study investigated the influence of a child's DRD4 risk, parental levels of ADHD symptoms, and the interactive influence of these factors on the development of preschool aggression. Additionally, the study investigated the role of home chaos as a mediator between parental ADHD symptoms and child aggression. The sample consisted of 84 4.5-year-old children and their parents. Children were genotyped for the DRD4 polymorphism. ADHD symptoms were self-reported by parents when the child was 2 to 6 months old. Parental reports of home chaos and the child's aggression were collected 4 years later. Child's DRD4 risk and parental ADHD symptoms significantly contributed to the prediction of preschool aggression. However, contrary to our hypotheses, no interactions were found between the child's DRD4 risk and the levels of parental ADHD symptoms. Home chaos played a mediating role in the relation between paternal ADHD symptoms and the child's aggression. The relation between maternal ADHD symptoms and the child's aggression was not significantly mediated through the level of home chaos. The current study emphasizes the importance of longitudinally investigating the contribution of parental ADHD symptoms to child aggression, while also exploring the differential contribution of maternal/paternal inattention and hyperactivity-impulsivity symptoms. Moreover, home chaos was found to be a significant environmental mechanism through which paternal ADHD symptoms affect children's aggression in the preschool years.


Online disease management of diabetes: engaging and motivating patients online with enhanced resources-diabetes (EMPOWER-D), a randomized controlled trial.
To evaluate an online disease management system supporting patients with uncontrolled type 2 diabetes. Engaging and Motivating Patients Online With Enhanced Resources for Diabetes was a 12-month parallel randomized controlled trial of 415 patients with type 2 diabetes with baseline glycosylated hemoglobin (A1C) values ≥7.5% from primary care sites sharing an electronic health record. The intervention included: (1) wirelessly uploaded home glucometer readings with graphical feedback; (2) comprehensive patient-specific diabetes summary status report; (3) nutrition and exercise logs; (4) insulin record; (5) online messaging with the patient's health team; (6) nurse care manager and dietitian providing advice and medication management; and (7) personalized text and video educational 'nuggets' dispensed electronically by the care team. A1C was the primary outcome variable. Compared with usual care (UC, n=189), patients in the intervention (INT, n=193) group had significantly reduced A1C at 6 months (-1.32% INT vs -0.66% UC; p<0.001). At 12 months, the differences were not significant (-1.14% INT vs -0.95% UC; p=0.133). In post hoc analysis, significantly more INT patients had improved diabetes control (>0.5% reduction in A1C) than UC patients at 12 months (69.9 (95% CI 63.2 to 76.5) vs 55.4 (95% CI 48.4 to 62.5); p=0.006). A nurse-led, multidisciplinary health team can manage a population of diabetic patients in an online disease management program. INT patients achieved greater decreases in A1C at 6 months than UC patients, but the differences were not sustained at 12 months. More INT than UC patients achieved improvement in A1C (>0.5% decrease). Trial registered in clinical trials.gov: #NCT00542204.


Surgical treatment of myopic strabismus fixus: a graded approach.
Surgical treatment of myopic strabismus fixus is challenging. Options for its correction range from conventional combined recession-resection surgery to innovative surgical procedures aiming to correct the deviated muscle paths. In this report we review our experience and compare the results of various surgical options for treatment of strabismus fixus. We report the surgical outcomes of nine adults with acquired strabismus fixus due to myopia with a follow-up of 1 year. Patients were enrolled between May 2003 and April 2007 in this retrospective study. The surgical procedure was determined depending on the angle of deviation and extent of motility impairment. A new transposition technique was performed in one patient who had an extreme variant of strabismus fixus. Combined recession-resection surgery was performed in four patients with resulting small-angle esotropia. In patients with both esotropia and hypotropia due to muscle alignment, we performed an additional upward displacement of both horizontal recti muscles combined with a myopexy of the lateral rectus muscle. The results were satisfying; in particular in one patient who had a transposition procedure a significant improvement was achieved. For treatment of myopic strabismus fixus, a graded approach seems advisable. Combined recession-resection surgery yields good results for smaller deviations with mildly impaired motility, additional fixation techniques need to be applied once the horizontal muscle paths are deviated, and in extreme cases, a transposition procedure is required.


Nature-based solutions for urban landscapes under post-industrialization and globalization: Barcelona versus Shanghai.
Using Barcelona and Shanghai as case studies, we examined the nature-based solutions (NBS) in urban settings-specifically within cities experiencing post-industrialization and globalization. Our specific research questions are: (1) What are the spatiotemporal changes in urban built-up land and green space in Barcelona and Shanghai? (2) What are the relationships between economic development, exemplified by post-industrialization, globalization, and urban green space? Urban land use and green space change were evaluated using data derived from a variety of sources, including satellite images, landscape matrix indicators, and a land conversion matrix. The relationships between economic development, globalization, and environmental quality were analyzed through partial least squares structural equation modeling based on secondary statistical data. Both Barcelona and Shanghai have undergone rapid urbanization, with urban expansion in Barcelona beginning in the 1960s-1970s and in Shanghai in the last decade. While Barcelona's urban green space and green space per capita began declining between the 1950s and 1990s, they increased slightly over the past two decades. Shanghai, however, has consistently and significantly improved urban green space and green space per capita over the past six decades, especially since the economic reform in 1978. Economic development has a direct and significant influence on urban green space for both cities and post-industrialization had served as the main driving force for urban landscape change in Barcelona and Shanghai. Based on secondary statistical and qualitative data from on-site observations and interviews with local experts, we highlighted the institution's role in NBS planning. Furthermore, aspiration to become a global or globalizing city motivated both cities to use NBS planning as a place-making tool to attract global investment, which is reflected in various governing policies and regulations. The cities' effort to achieve a higher status in the global city hierarchy may have contributed to the increase in total green space and urban green per capita. In addition, various institutional shifts, such as land property rights in a market economy vs. a transitional economy, may also have contributed to the differences in efficiency when expanding urban green space in Barcelona and Shanghai.


Signal enhancement and suppression during visual-spatial selective attention.
Selective attention involves the relative enhancement of relevant versus irrelevant stimuli. However, whether this relative enhancement involves primarily enhancement of attended stimuli, or suppression of irrelevant stimuli, remains controversial. Moreover, if both enhancement and suppression are involved, whether they result from a single mechanism or separate mechanisms during attentional control or selection is not known. In two experiments using a spatial cuing paradigm with task-relevant targets and irrelevant distractors, target, and distractor processing was examined as a function of distractor expectancy. Additionally, in the second study the interaction of perceptual load and distractor expectancy was explored. In both experiments, distractors were either validly cued (70%) or invalidly cued (30%) in order to examine the effects of distractor expectancy on attentional control as well as target and distractor processing. The effects of distractor expectancy were assessed using event-related potentials recorded during the cue-to-target period (preparatory attention) and in response to the task-relevant target stimuli (selective stimulus processing). Analyses of distractor-present displays (anticipated versus unanticipated), showed modulations in brain activity during both the preparatory period and during target processing. The pattern of brain responses suggest both facilitation of attended targets and suppression of unattended distractors. These findings provide evidence for a two-process model of visual-spatial selective attention, where one mechanism (facilitation) influences relevant stimuli and another (suppression) acts to filter distracting stimuli.


Molecular evolution of broadly neutralizing Llama antibodies to the CD4-binding site of HIV-1.
To date, no immunization of humans or animals has elicited broadly neutralizing sera able to prevent HIV-1 transmission; however, elicitation of broad and potent heavy chain only antibodies (HCAb) has previously been reported in llamas. In this study, the anti-HIV immune responses in immunized llamas were studied via deep sequencing analysis using broadly neutralizing monoclonal HCAbs as a guides. Distinct neutralizing antibody lineages were identified in each animal, including two defined by novel antibodies (as variable regions called VHH) identified by robotic screening of over 6000 clones. The combined application of five VHH against viruses from clades A, B, C and CRF_AG resulted in neutralization as potent as any of the VHH individually and a predicted 100% coverage with a median IC50 of 0.17 µg/ml for the panel of 60 viruses tested. Molecular analysis of the VHH repertoires of two sets of immunized animals showed that each neutralizing lineage was only observed following immunization, demonstrating that they were elicited de novo. Our results show that immunization can induce potent and broadly neutralizing antibodies in llamas with features similar to human antibodies and provide a framework to analyze the effectiveness of immunization protocols.


Circulating MicroRNAs as Prognostic Molecular Biomarkers in Human Head and Neck Cancer: A Systematic Review and Meta-Analysis.
Circulating microRNAs (miRNAs) are potential molecular biomarkers for cancer detection; however, little is known about their prognostic role in head and neck cancer. This current study is aimed at evaluating the role of novel miRNAs in the survival of head and neck cancer patients. We performed a systematic literature search using online databases for articles published between December 2006 and February 2019. A meta-analysis was conducted to assess the correlation between miRNA expressions and overall survival (OS) among the selected head and neck cancer studies. After multilevel screening by reviewers, meta-analysis was performed using hazard ratios (HR) and associated 95% confidence interval (CI) of survival to calculate a pooled effect size. A total of 1577 patients across 13 studies were included in the literature review, with 18 miRNAs upregulated and 4 miRNAs downregulated predicting a poor overall survival. The forest plot generated using cumulated survival data resulted in a pooled HR value of 2.943 (95% CI: 2.394-3.618) indicating a strong association of dysregulated miRNA expression with a poor outcome. Only 2 miRNAs-low levels of miR-9 and high levels of miR-483-5p-were observed in two studies, both showing a significant association with overall cancer survival. To our knowledge, this is the first comprehensive systematic review and meta-analysis that examines the prognostic role of circulating miRNAs from blood in head and neck cancer patients. The combined effect estimates a HR across multiple studies and also supports the previous individual findings that an alteration in miRNA expression is highly associated with poor prognosis. This has the potential to use serum and/or plasma miRNAs as biomarkers and become novel tools for predicting the prognosis of head and neck cancer patients in the near future.


CTLA-4 control over Foxp3+ regulatory T cell function.
Naturally occurring Foxp3+CD4+ regulatory T cells (Tregs) are essential for maintaining immunological self-tolerance and immune homeostasis. Here, we show that a specific deficiency of cytotoxic T lymphocyte antigen 4 (CTLA-4) in Tregs results in spontaneous development of systemic lymphoproliferation, fatal T cell-mediated autoimmune disease, and hyperproduction of immunoglobulin E in mice, and it also produces potent tumor immunity. Treg-specific CTLA-4 deficiency impairs in vivo and in vitro suppressive function of Tregs-in particular, Treg-mediated down-regulation of CD80 and CD86 expression on dendritic cells. Thus, natural Tregs may critically require CTLA-4 to suppress immune responses by affecting the potency of antigen-presenting cells to activate other T cells.